Anti-GCLM antibody [EPR6667]

6 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cardiac muscle tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Western blot - Anti-GCLM antibody [EPR6667] (ab126704)

  All lanes: Western blot - Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

  Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

  Lane 3: PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 31 kDa

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  Western blot - Anti-GCLM antibody [EPR6667] (ab126704)

  Lanes 1 - 3: Merged signal (red and green). Green - ab126704 observed at 31 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
  
  ab126704 was shown to recognize GCLM in wild-type HAP1 cells as signal was lost at the expected MW in GCLM knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GCLM knockout samples were subjected to SDS-PAGE. ab126704 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-GCLM antibody [EPR6667] (ab126704) at 1 µg/mL

  Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

  Lane 2: GCLM knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HeLa whole cell lysate at 20 µg

  Predicted band size: 31 kDa

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  Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)

  Purified ab126704 at 1/20 dilution (0.6μg) immunoprecipitating GCLM in HeLa whole cell lysate.
  Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
  Lane 2 (+): ab126704 + HeLa whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab126704 in HeLa whole cell lysate.
  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/5000 dilution) was used for Western blotting.
  Blocking Buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM/TBST.
  Observed band size: 31 kDa

  All lanes: Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)

  Predicted band size: 31 kDa